Penicillin compositions



Patented July 26, 1949 STATE .ZPENICIL'LIN CGMPOSITIONS John OLBower, Wyncote, Pa.

Nn'i'Drawing; Application December 26,1944; Serial No. 569,919

4 Claims. 1

This inventionsrelates stosmethods and :compositions for prolonging; themeffectiueness Y of therapeutic :and medicinalsagents, and for making; it 'possiblesto produce. a given. curative and bactericidal efiect by the usehof a smaller quantity of the agent employed; Although the invention is -:applicab1e -.to various. therapeutic and antiseptic :wagents it is: particularly valuable in'connectloncw-ith penicillimand for this reason penicillin will be referred-to .in this specification as illustrating the invention;

It has long been recognized-that the body tends to resent the presence of alien substances, even the most salutary-remedies, and that it acts to isolate or throw offz any: substance not: of its own making. isldramatically, illustrated in the case 50f: penicillin; Treatment with this remedy requires-the administration soflrelat-ivelylarge quantities at relatively frequent intervals. Almost -60 -p'er roent nfthe amount of.- onenicillinintroduced intravenously; ;for .example, is thrown ofl- :by the :kidneys .-in:' thevurine :withinethe first hour folloWingJts infraction;

I have discovered vthat if thezpenicillin. is introduced in the .form of .a :special esolution incorp orating constituents-otanimal blood (which-solution is more pantioulamlmdescri-b'ed hereinafter) the penicill-inzwill- .-remain:.:in=r the a-body when injected into the eblood- -.or muscle 1 :or :-other tissue or placed in .-a?cavity;.-of the-body; tor; instance, the peritonealcavity,- ion-a much longer interval. I have also :QbSfilVQd that it is absorbed-more slowly iirom thelsurtace lotthelbody Whether the skin belintact or partially or completely devitalized as .in the [case-of burnsor iii-diseased offanimahblood which are entirely comparable with those obtained by others who, lacking knowledge of theinvention, have found it'necessary to administer considerably; larger quantities of penicillin and havefoundiit'necessary. to re peat the treatmentlatimu'ch more frequent intervals.

The penicillin is, as indicated above,admin-istered' in the form of. an admixture, the other component of which comprises. constituents of animal blood. In preparing such. an admixture for the treatment ofburns and other external wounds, I. prefer 'to employ. about. 400 to 500 units of penicillin @tos'eaeh @cublocentimeter of blood constituents.- For -'':intravenous injection, from 400 to 1,000 units :pereccl may be employed,

As a practical matter, as little as 200 units of.

penicillin per co. produces sa marked healing effect; while, on the-other :hand;- drastic situations may callaforsthe-euse of asmuch -as--3,000 units per cc.v The erangcofwconcentration is great, since penicillin .ls: quite? readily soluble in the solution herem-desorlbed,

Animal blood may be regarded as containing four-major constituents;.red andwhite blood cells, blood platelets and-plasma; An essential characteristic of i my inyentioniis .-that the prep aration contains as roneecomponent Of'Tthe Jadmixture the stroma,'-orstructural *element, of the red blood cell, ins-3a. state-pfisubstantially complete physical disintegratiomz. A preferred aspect of the ;-invention;dealsr withiseparation of the plasma before .adding th'e penicillin; to the remaining constituent-sh When ithisgafis 'done, plasma may subsequently mbe addedoito: the material in any desired 'amountggor-irthe material may be used in ,aisubstantially plasmae'free. cone dition. i

In my; copen'ding: applications, Serial No. 502,503, filed Septemberd-d;1'9't3;. and-Serial No. 519,686, filed January 25.;5 I944, amew form-cf blood cell solution; containing-hemoglobin, electrolytes and disintegrated stroma, is disclosed. That solutionis particularly advantageous as the other component for vthe penicillin admixture of the present invention; application-s contain rather full instructions flier preparing blood solutions having-thecharacteristic abovereferred to, which instructions will. be only summerized herein.

Briefly, these solutions arapreparedior preference, from those componentsof the blood which remain afteryextraction of-the plasma. The extraction is readily efiectedafter' centrifuging, by vacuum suction-into-asterile ampoule, all but a very small amount of the-blood plasma being removed. The remainingblood resi'due, consisting essentially of red-andwhite blood .cells, blood platelets, electrolytes-and'gthe' small amount of plasma, is then subjected to anlextremely rapid and deep-freezing'operation. A distin uishing feature of this process is therapiditywith which the mixture or'residue is frozen.v In, practice; this extremely rapld'jfreezing actionis most readily secured byimm'ersing a vessel containing the blood constituents in 1a mixture of, Dry Ice and Sunoco? spirits, this combinationafiording temperatures intheneighborhood.v of C. or C. (Sunocoi spirits .is .alrefinedkerosene.) The intense-v cold so produced willcome pletely disintegrate the blood'cellstructure in not more than-th'ree minutes itthe solution is in a 30 cc. test tube, and may produce this effect in as short a time as sixty seconds. Results at lea-st equally good may be obtained in as little as seven or eight seconds if a vessel of the same size is immersed in liquid air or in liquid nitrogen; or in a proportionately longer interval if larger vessels are used. However, these lastnamed refrigerants are somewhat difiicult to handle-primarily because of the danger of breaking ordinary laboratory glassware by subjecting it suddenly to such drastic temperature changes. Liquid air, for example, manifests temperatures of the order of 192" C. This breakage can be obviated by the use of stainless steel or similar unbreakable containers. It should also be remembered that liquid air is inflammable, and, therefore, somewhat more dangerous than either liquid nitrogen or solidified carbon di oxide. v

The result of this rapid and deep freezing is to completely disrupt the structure of the blood cell-s and platelets. I have used the term disintegrate to describe this phenomenon, which is quite different from the customary breakdown of blood cell structure produced by mild freezing or even without freezing by the addition of distilled water or chemicals. The distintegration produced by the rapid freezing just described reduces the structure of the blood cells so thoroughly that no differentiation between the blood cell liquid (hemoglobin and electrolyte) and the stroma has been observed even under microscopic examination with either the 4 mm. or the 16 mm. lenses. The solution produced is filterable. In fact it has on many occasions been passed through the asbestos pad of the Horm bacteriological filter now commonly employed to remove micro-organisms from contaminated plasma. Pads of this type are known to be more effective than the finest filter papers, and it is astonishing that any such unfilterable material as red blood residue. could be so treated as to meet this test. Yet the fact has been verified by chemical analysis and by spectroscopic examination.

It may be that the solution. produced is truly colloidal in character. On the other hand, it may even be true that the change effected in the nature of the-stroma is such as to render this substance literally soluble, to a greater or lesser degree, in the blood cell content. Since I have been unable to establish with certainty whether the stroma is actually dissolved in the hemoglobin-electrolyte mixture, or whether it is present therein in a colloidal or subcolloidal form, the expression solution is used with the intention of embracing either alternative.

Obviously, the actual time required for deepfreezing a given quantum of material will vary, according to the surface area of the vessel in proportion to its volume, the temperature of the coolant, the rate of heat-transfer through the vessel walls, etc. No conventional standard is available as a guide for determining when a sufficiently rapid rate of freezing has been attained. Under the circumstances, it is necessary to use the results of the Dry-Ice method as a standard of reference in defining the nature of the freezing step: that is, as noted above, my invention contemplates eifecting a degree of disintegration in the structure of blood cells and platelets equivalent to that produced by immersing a 30 cc. test tube containing a substantially plasma-free blood solution in a mixture of Dry Ice and Sunoco spirits at a temperature of from -'I0 C. to

4 C. for an interval of from one to three minutes.

In the two applications mentioned above, I have disclosedmore fully the nature of the blood solution described in the foregoing paragraphs, and have suggested several ways in which it can be used. In my copending application, Serial No. 547,318, filed July 29, 1944, there is described a method for desiccating such solutions, containing the hemoglobin, electrolyte and disintegrated stroma, and there are offered suggestions as to ways in which the dry, amorphous powder so produced may be employed. The technique followed in producing this dry powder is analogous to that employed in lyophilizing plasma, and involves gradually desiccating the hemoglobin-electrolyte-stroma solution under controlled temperature and vacuum conditions, so as to leave an amorphous residue which is substantially free of moisture.

An important characteristic of the products obtained by all of the quick-freezing techniques discussed is an astonishing resistance to change during storage. The products, whether in powder or in liquid form, will keep without deterioration for much longer intervals than animal blood which has not been subjected to the deep-freeze treatment.

According to the present invention, there are several methods for obtaining the special admixture of penicillin and blood constituents, all involving the employment of blood constituents derived by applying a quick-freezing treatment to human blood.

First, as the preferred method, I freeze and then thaw the blood residue, as described herein, and after it has completely melted, the desired amount of penicillin is added. This admixture is then drawn into a sterile vacuole and re-frozen by a process called shelling, necessary to proper desiccation. shelling prepares the material for drying efficiently. Thevacuole is placed horizontally in a bath of Dry Ice and Sunooo spirits and revolved on its axis continuously at an even rate, the solution forming a thin shell on the sides of the ampoule and having a hollow in the middle. The ampoule is then connected to an intake manifold by means of a rubber nipple. The manifold is attached to a condenser with a secondary coil which is in turn attached to a vacuum pump. As the frozen liquid volatilizes it is drawn off and condensed. After 48 hours the ampoule containing the fine dry powder is removed and stoppered under vacuum, using sterile technique.

The resulting preparation may be placed in condition for intravenous injection, for example, by simply adding plasma, or by adding plasma and distilled water, or by adding distilled water alone. The nature of the addition will influence the viscosity of the liquid produced. For external application, the dry powder may simply be sprinkled on the Wound, or it may be mixed with plasma and/or water and applied as a liquid or as a paste.

The second method for constituting the admixture is to add the desired amount of penicillin (in aqueous solution for convenience) directly to the hemoglobin-electrolyte-strcma solution, without desiccating that solution. This procedure is advantageous where the product is to be used within a relatively short time-that is, within a matter of several days. This is the procedure usually employed when the solution is used locally, as, for instance, in the treatment of burns. It is frequently advantageous to add plasma or "plasma; Incidentally, the admixtures referred to may be diluted with normal saline solution to any extent desired. Unusually rapid healing and asubstantial reduction in pain follows the application to burns and external Wounds of the blood-constituent solution admixed with the mixtures of alkyl dimethyl benzyl ammonium chlorides more fully described in New and Non-officialRemedies, edition of 1944, at page 112. This mixture is commercially available under the trade-mark Zephiran Chloride, or cetylpyridinium chloride, a quaternary ammonium compound, known under the trade-mark Ceepryn; so also where certain of the sulphonamides, particularly sodium sulphathiazole, have been used in forming the admixture.

In short, an admixture of the hemoglobin-electrolyte-stroma solution with each of a number of remedial agents has resulted in an enhancement of the curative and bactericidal eifect of the agent used. From the information given it will be apparent to those skilled in the healing arts that a similar improvement in the effectiveness of other healing agents is to be expected upon admixing them with this special blood solution. It will also be obvious that agents which manifest an unfavorable efiect upon blood-that is, which are by nature incompatible with human blood, should not be employed. However, very few compounds not compatible with blood are regarded as remedial agentsthat is, as belonging to the class of therapeutic materials with which my invention is especially concerned.

Another aspect of thisrinvention, taking advantage of the surprising resistance of these compounds to deterioration, is the possibility of incorporating them in ointments. Various. ointment-base formulae have been tried. My present preference is for one incorporating a simple foundation (such as a cerate and/or petroleum jelly) and zinc peroxide, boric acid, and zinc oxide. To such a base, 10% by weight of the hemoglobinelectrolyte-stroma solution (or a corresponding proportion of the desiccated material) is added, in admixture with 600 units of penicillin per 00. The ointment so produced has been applied to lesions of the epidermis as well as the deeper tissues, and healing has uniformly taken place thereafter at a maximum rate. This has been especially marked where the ointment has been used in the treatment of burns, and the more extensive the damage, the more dramatic is the recovery.

Wherever the preparations described herein have been used, I have been impressed with an apparent increase in the body tolerance towards the curative agent employed, and an apparent improvement in the rapidity and certainty of healing. The improvement in a patients-condition, following an intravenous injection of this special red blood material admixed with penicillin does not wear off in the space of a couple of hours, as it does where penicillin alone is used. An almost immediate reduction in pain follows the application of the admixture (especially in ointment form) to burns of any degree, and this effect persists for several hours even in themost serious cases. 1

There is a definite gain in the treatment which I have developed simply from the fact that the patient need be disturbed less frequently. Add to this the saving in vitality which results from the fact that, because of the decrease in the quantity required, the body expends less efiort in throwing ofi the drug. And if, as I suspect, the form in,

which the material is administered results in an enhancement in its efiect which is more than a function of the time it remains in the system, there is a great gain in this aspect of the case. It will be seenthat economy resulting from the reduction in amount required is but a small factor compared to the other benefits accruing from the present invention.

In conclusion it may be noted that, although the foregoing discussion specifically relates to the treatment of blood which has been citrated (this being standard practice in the taking of blood from donors), it is possible, and under some circumstances may be highly desirable, to subject the pure blood to the treatment which I have invented. This can be safely accomplished if the blood withdrawn from the donor is deep-frozen promptly. The hemoglobin-electrolyte-stroma solution prepared in this way has, according to laboratory investigation, shown a higher proportion of hemoglobin than is found in whole blood to which sodium citrate has been added. It may possess attributes of considerable importance, since it is free of any extraneous material whatsoever, and may, therefore, have an especial usefulness in therapeutic work.

Where disintegration is referred to in the claims, it is to be understood that the term is used to define the condition discussed heretofore, namely, so complete a disruption of the red blood cells that discrete particles of their structure (stroma) cannot be differentiated under examination with the 4 mm. or 16 mm. lenses of an ordinary lab-oratory microscope.

I claim:

l. A solution for therapeutic use incorporating penicillin and the constituents of animal blood remaining after centrifugal extraction of plasma, the preparation being characterized in that the blood stroma which it contains is present in a state of dissolution such that it cannot be visually differentiated from the remaining blood constituents under microscopic examination under the 4 mm. lens.

2. An ointment for clinical administration incorporating about by weight of an ointment base and about 10% by weight of the solution of claim 1.

3. The impalpable powder resulting from vacuum desiccation of the solution of claim 1.

4. An ointment for clinical administration containing an ointment base and the solution of claim 1. 1

JOHN O. BOWER.

REFERENCES CITED The following referenlces are of record in the file of this patent:

Comptes Rendus de la Soc. de Biol. (1900), page 

